Dynabeads

The physicians used real-time X-ray technology to visualize the patient’s vascular system and locate IA inside the blood vessel. A solitaire stent retriever (Covidien, Irvine, CA, United States) was used for biopsy specimens’ retrieval from an occluded IA. Cerebral aneurysm and malformed arteries were isolated from cerebral arties using a cutting plane and separating the surfaces on either side of the plane by using a minimally invasive technique. Endothelial cells were derived from human cerebral aneurysm and malformed arteries, and separated with immunomagnetic cell sorting. Briefly, Anti-CD 146-coated Dynabeads (Invitrogen, CA, United States) were prepared according to manufacturer’s instruction and stored at 4°C. Fifty-mg aneurysms were ground with glass pestle and mortar with one-millilter PBS buffer, 0.1% bovine serum albumin, 0.1% sodium azide, and 0.1% a standard broad-spectrum inhibitor cocktail at 4°C. Ten-microliter FcR-blocking agent (Miltenyi, Bergisch Gladbach, Germany) and 25-microliter antibody-coated Dynabeads were added and mixed thoroughly. The samples were mixed in a mixer for 1 h at 4°C and washed four times with PBS inside the Big Easy Magnet (EasySep, United States) at 4°C. Between each washing procedure, the endothelial cells were flushed out with MACS buffer (PBS with 0.5% BSA and 2 mM EDTA, pH 7.0) 10 times in a 100-μL pipette.